CIHM 
Microfiche 
Series 
(IMonographs) 


ICIMH 

Collection  de 
microfiches 
(monographies) 


Canadian  Instituta  for  Historical  Microraproductions  /  Institut  Canadian  da  microraproductions  historiquas 


Technical  and  Bibliographic  Notes  /  Notes  techniques  et  bibliographiques 


The  Institute  has  attempted  to  obtain  the  best  original 
copy  available  for  filming.  Features  of  this  copy  which 
may  be  bibliographicaily  unique,  which  may  alter  any  of 
the  images  in  the  reproduction,  or  which  may 
significantly  change  the  usual  method  of  filming  are 
checked  below. 

n^  Coloured  covers  / 
I — I  Couverture  de  couleur 


D 
D 


D 


Covers  damaged  / 
Couverture  endommag^e 

Covers  restored  and/or  laminated  / 
Couverture  restaur^  et/ou  pellicul^e 

Cover  title  missing  /  Le  titre  de  couverture  manque 

Coloured  maps  /  Cartes  gSographiques  en  couleur 

Coloured  ink  (i.e.  other  than  blue  or  black)  / 
Encre  de  couleur  (i.e.  autre  que  bieue  ou  noire) 


I      I   Coloured  plates  and/or  illustrations  / 


D 
D 

a 


n 


Planches  et/ou  illustrations  en  couleur 

Bound  with  other  material  / 
ReliS  avec  d'autres  documents 

Only  edition  available  / 
Seule  Edition  disponible 

Tight  binding  may  cause  shadows  or  distortion  along 
interior  margin  /  La  reliure  serr^e  peut  causer  de 
I'ombre  ou  de  la  distorsion  le  long  de  la  marge 
int^rieure. 

Blank  leaves  added  during  restorations  may  appear 
within  the  text.  Whenever  possible,  these  have  been 
omitted  from  filming  /  Use  peut  que  certaines  pages 
blanches  ajout^es  lors  d'une  restauration 
apparaissent  dans  le  texte,  mais,  iorsque  ceia  ^tait 
possible,  ces  pages  n'ont  pas  i\6  fiim^es. 


a 


L'Institut  a  microfilm^  le  meilleur  exemplaire  qu'il  lui  a 
6X6  possible  de  se  procurer.  Les  details  de  cet  exem- 
plaire qui  sont  peut-6tre  uniques  du  point  de  vue  bibli* 
ographique,  qui  peuvent  modifier  une  image  reproduite. 
ou  qui  peuvent  exiger  une  modification  dans  la  m^tho- 
de  normale  de  filmage  sont  indiqu^s  ci-dessous. 

I     I  Coloured  pages  /  Pages  de  couleur 

I I  Pages  damaged  /  Pages  endommag6es 


D 


Pages  restored  and/or  laminated  / 
Pages  restaurdes  et/ou  pellicul^es 


0  Pages  discoloured,  stained  or  foxed  / 
Pages  d^color^es,  tachet^es  ou  piqu^es 

I     I  Pages  detached  /  Pages  d6tach6es 

I  /}  Showthrough / Transparence 

I      I  Quality  of  print  varies  / 


□ 


D 


Quality  in^gale  de  I'impression 

Includes  supplementary  material  / 
Comprend  du  materiel  suppl^mentaire 

Pages  wholly  or  partially  obscured  by  errata  slips, 
tissues,  etc.,  have  been  refilmed  to  ensure  the  best 
possible  image  /  Les  pages  totalement  ou 
partiellement  obscurcies  par  un  feuillet  d'errata,  une 
pelure,  etc.,  ont  6\6  film  °s  k  nouveau  de  fa^on  k 
obtenir  la  meilleure  image  possible. 

Opposing  pages  with  varying  colouration  or 
discolourations  are  filmed  twice  to  ensure  the  best 
possible  image  /  Les  pages  s'opposant  ayant  des 
colorations  variables  ou  des  decolorations  sont 
film^es  deux  fois  afin  d'obtenir  la  meilleure  image, 
possible. 


Additional  comments  / 
Commentaires  suppl6mentaires: 


Pagination  is  as  follows:     p.  329-3%%. 


This  It 
Cedo 

lOx 

em  is  filmed  at  the  reduction  ratio  checked  below  / 
Bument  est  filme  au  taux  de  reduction  indiqui  ci-dessout. 

14x                               18x 

22x 

26x 

30x 

>/ 

12x 

16x 

20x 

24x 

28x 

32x 

J 

Th«  copy  filmtd  h«r«  has  b««n  raproductd  thanks 
to  tha  ganarosity  of: 

Blacker-Wood  Library  of  Biology 
McCill   University,  Montreal 


L'axamplaira  film4  fut  raproduit  grica  *  la 
gAnArosit*  da: 

Blacker-Wood  Library  of  Biology 
HcCin  University,  Montreal 


Tha  imagas  appaaring  hara  ara  tha  bast  quality 
possibia  considaring  tha  condition  and  lagibility 
of  tha  original  copy  and  in  kaaping  with  tha 
filming  contract  apacificationa. 


Las  imagas  suivantas  ont  ttt  raproduitas  avac  la 
plus  grand  soin.  compta  tanu  da  la  condition  st 
da  la  nattatA  da  l'axamplaira  film*,  at  an 
conformity  avac  las  conditions  du  contrat  da 
filmaga. 


Original  copies  in  printod  papar  eovars  ara  fllmad 
beginning  with  tha  front  covar  and  anding  on 
tha  last  paga  with  a  printad  or  iliustratad  impraa- 
sion,  or  tha  back  covar  whan  appropriate.  All 
othar  original  copies  are  filmed  beginning  on  the 
first  page  with  a  printed  or  illuatratad  imprea- 
sion.  and  anding  on  the  laat  paga  with  a  printed 
or  illuatratad  impreasion. 


Las  axemplaires  originaux  dont  la  couvartura  an 
papier  eat  imprimia  sont  filmis  an  commandant 
par  la  premier  plat  at  en  terminant  soit  par  la 
derniire  paga  qui  comporta  une  emprainta 
d'impression  ou  d'illustration.  soit  par  la  second 
plat,  salon  le  cas.  Tous  las  autres  axemplaires 
originaux  sont  filmis  en  commen^ant  par  la 
pramiire  page  qui  comporte  une  empreinte 
d'impreasion  ou  d'illustration  at  en  terminant  par 
la  darniire  paga  qui  comporte  une  telle 
empreinte. 


The  last  recorded  frame  on  eech  microfiche 
shall  contain  tha  symbol  — *•  (meaning  "CON- 
TINUED"), or  the  symbol  ▼  (meaning  "END"), 
whichever  applies. 


Un  des  symboles  suivants  Apparaitra  sur  la 
darniire  image  do  cheque  microfiche,  salon  le 
ces:  le  symbols  — ^  signifie  "A  SUIVRE".  le 
symbols  ▼  signifie  "FIN  ". 


Maps,  plates,  charts,  etc.,  mey  be  filmed  at 
different  reduction  retios.  Those  too  large  to  be 
entirely  included  in  one  exposure  ara  filmed 
beginning  in  the  upper  left  hend  corner,  left  to 
right  and  top  to  bottom,  as  many  frames  as 
required.  The  following  diagrams  illustrate  the 
method: 


Lea  certes,  planches,  tableaux,  etc..  peuvant  itre 
filmte  i  des  taux  da  reduction  diff Grants. 
Lorsqua  le  document  est  trop  grand  pour  Atre 
raproduit  an  un  seul  cliche,  il  est  film*  A  partir 
da  Tangle  supirieur  gauche,  de  gauche  A  droite. 
et  de  haut  en  bes.  en  prenant  le  nombre 
d'imeges  nicessaira.  Um  diagrammes  suivants 
illustrant  la  mtthoda. 


1  2  3 


1  2  3 

4  5  6 


MicKocory  resoiution  mt  chaut 

(ANSI  and  ISO  TEST  CHART  No.  2) 


1^  IZ-B 

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^  /APPLIED  IIVHGE    I 

^K  1653  East   Uoin  Street 

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^"^  WS^^^LI  \LI  RESERVE  OF  MARINE  FISH  AND       o    \  \ 
JBRARY  INVERTEBRATES  ^^ 


THfe^XCRETION  OF  CARBON  DIOXIDE 


C\f/ 


BY 

J.  B.  COLLIP 


(From  the  Marine  Biological  Station,  Departure  Bat,  Canada) 


<;f' 


RiPBiinno  FBOH 

THE  JOURNAL  OF  BIOLOGICAL  CHEMISTRY 
Vol.  XLIV,  No.  2,  November,  1920 


ffK^^MX^imittttlBB^II^ 


Raprinted  from  Ta*  Journai.  or  Biolouical  Cbemivtiit,  Vol.  XLIV,  No.  2,  November,  1920 


THE  ALKALI  RESERVE  OF  MARINE  FISH  AND 
INVERTEBRATES. 

THE  EXCRETION  OF  CARBON  DIOXIDE. 

Bt  J.  B.  COLLI  P. 
{From  the  Marine  Biological  Station,  Departure  Bay,  Canada.) 

(Received  for  publication,  September  3, 1020.) 
INTRODUCTION. 

The  results  of  an  investigation  to  determine  the  carbon  dioxide 
content  of  the  blood  and  body  fluids  of  such  marine  forms  as 
were  available  for  study  at  or  in  the  vicinity  of  the  Marine  Biolog- 
ical Station,  Departure  Bay,  Vancouver  Island,  British  Columbia, 
are  herein  reported.  As  the  Van  Slyke-Cullen  (1)  apparatus 
furnishes  a  convenient  and  ready  means  of  determining  the  car- 
bon dioxide  content  and  capacity  of  blood  and  thereby  the  alkali 
reserve  of  the  same,  it  was  used  exclusively  in  this  investigation. 

Methods. 

The  representd,tive  forms  studied  were,  for  the  most  part,  col- 
lected personally.  Great  care  was  taken  to  insure  that  the  indi- 
vidual specimens,  the  blood  or  celomic  fluid  of  which  was  to  be 
examined,  should  be  bled  while  in  a  fresh  condition.  As  will 
appear  more  fully  in  a  subsequent  communication  this  is  a  very 
essential  point  especially  as  regards  various  molluscan  types. 
An  endeavor  was  made  to  secure  specimens  representative  of  as 
many  of  the  invertebrate  phyla  and  orders  of  the  Pisces  as  was 
possible. 

The  methods  employed  in  obtaining  blood  or  celomic  fluid 
from  the  forms  studied  will  now  be  detailed. 


32B 


330 


Alkali  Reserve 


Cmlenterata. 

Two  typcH  of  MedusjB  and  the  sand  anomonc  were  examined. 
The  results  recorded  for  these  specimens  can  only  be  taken  as 
approximate  since  the  methods  here  adopted  were  of  a  special 
character. 

Medus<r. — Several  specimens  were  caught  in  a  dip  net,  trans- 
ferred to  a  glass  container  filled  with  fresh  sea  wpter,  and  carried 
at  once  to  the  laborator>'.  Thoy  wore  first  drained  free  of  sea 
water  hy  siispendinR  ovor  a  wido  funnel  l>y  means  of  a  double 
fold  of  checsr-cloth.  Tliey  wcrr  llioii  Konlly  miicerated  and 
p;isw»(l  llintUKli  the  chcpsi'-clotli.  The  jelly-like  mass  was  col- 
lected ill  ii  clean,  dry  test-tulM-.  TIum  was  shaken  vigorously  for 
'.i  minutes,  air  being  admitted  on  several  occasions.  I  cc.  of  the 
fluid  was  then  transferred  to  the  Van  Slyke-Cullon  apparatus  and 
the  analysis  made  in  the  usual  manner.  The  evolved  carbon 
dioxide  was  absorbed  by  the  use  of  10  per  cent  sodium  hydroxide. 
The  volume  of  gas  was  reduced  to  cubic  centimeters  of  carbon 
dioxide  at  0°C.  and  760  mm.  pressure  held  by  100  cc.  of  fluid. 

Owing  to  the  difficulties  in  the  way  of  collecting  blood  or 
celomic  fluid  from  marine  forms  without  loss  of  carbon  dioxide  it 
was  thought  best  to  examine  all  specimens  under  uniform  con- 
ditions. The  various  samples  were  therefore  shaken  for  3  min- 
utes in  the  test-tulx"  or  25  cc.  Luer  syringe  with  atmospheric 
air  wliich  was  renewed  on  several  occasions.  The  samples  thus 
equilibrated  with  atmospheric  air  were  submitted  at  once  to 
analysis.  In  many  instances  concurrent  samples  were  also 
ecjuilibiuted  with  alveolar  air  of  the  normal  human  subjfect  after 
ihv  manner  suggested  by  \'an  Slyke  and  Cullen  (2).  Such 
sp(icimens  were  tlicMi  transferred  by  means  of  an  Ostwjdd  pipette 
to  the  apparatus  for  analysis  without  appreciable  loss  of  carbon 
dioxide. 

.SV(i  Aneniont.s.  The  s|}ecinien  was  dug  from  a  sand  flat  at 
low  water  and  carried  to  the  laboratory  in  a  container  filled  with 
fresh  sea  water.  The  animal  was  then  allowed  to  contract  in  a 
dry  open  vessel.  A  slit  was  next  made  in  the  side  by  means  of  a 
sharp  scalpel  and  the  cavity  drained  free  of  the  fluid.  The  soft 
parts  were  macerated  in  a  mortar  and  finally  .suspended  in  an 
equal  volume  of  fresh  distilled  water.    2  cc.  of  the  suspension 


t^^j^jk 


J.  B.  CoUip 


331 


were  analyzed  and  the  results  interpreted  as  being  approximately 
indicative  of  the  carbon  dioxide  content  of  the  tiHsues  of  the 
animal. 

JirachioiHMlit. 

The  specimens  were  collected  at  low  water  from  the  rocks 
closely  adjacent  to  the  lalwratorv  and  examined  immediately. 
It  was  found  that  from  1  to  2  cc.  of  fluid  could  be  obtained  from 
these  small  forms  by  aspirating  into  a  Luor  syringe  from  the  celomic 
cavity  through  a  needle  passed  through  the  soft  tissue  <!xpo.sed  at 
the  hinge. 

Echinotlrnnatn. 

Starfish.  'VhcHo  were  collected  near  the  laboratory  at  low 
water  and  examined  inuiiediutely.  The  colomie  fluid  was  aspi- 
rated from  the  celomic  pouch  of  one  of  the  arms.  This  was 
entered  dorsally  and  laterally. 

Sea  I 'rch ins.  The  celomic  fluid  was  aspirated  from  a  celomic 
pouch  in  much  the  same  manner  as  in  the  case  of  the  starfish. 

Arthropoda. 

Crufitacea.  Several  sjx'cies  of  Crustacea  were  studied.  Blof»d 
was  obtained  frotii  the  individual  specimens  by  introducing  :i 
hypodermic  needle  into  the  pericardium  and  aspirating  into  a 
liUer  syringe.  In  the  case  "f  the  larger  forms  such  as  Hchiil- 
nocerus  formatus  and  Canar  magistir  a  small  hole  was  first 
trephined  through  the  carapace  just  dorsal  to  the  pericurdiuin 
The  needle  was  then  introduced  through  this  opening.  It  w.-is 
found  that  the  needle  could  be  pas.sed  directly  through  '  ,• 
carapace  of  small  crabs  and  shrimps.  Blood  was  obtainetl  fr<»ni 
the  pericardial  sinus  of  the  eagle  barnacle  {Jialanu.^  tuiuilln)  aftrr 
a  .small  hole  had  been  drilled  through  the  calcareous  shell  just 
dorsal  to  the  sinus. 

MoUuacn. 

Pelecypodn. — Many  fonns  representative  of  different  si)e('ies 
of  the  pelecypod  Mollusca  were  examined.  The  celomic  fluid  or 
"clam  juice"  was  obtained  in  all  instances  by  aspirating  from  the 


te- 


9 
f- 


332 


Alkali  Reserve 


pericardiul  cavity.  The  needle  was  introduced  into  the  sinus 
throuRh  the  soft  tissue  in  the  mid-line  dorsal  to  the  latter. 

GuKlropoda. — Four  different  species  were  obta'ncd  for  study. 
Celoniic  fluid  was  obtained  from  the  pericardial  Siuus  of  the  two 
jihcll  forms  Polynices  (Lunatia)  Inrini  and  Thais  lamellom, 
after  a  small  opening  had  been  drilled  in  the  shell  adjacent  to  the 
sinus.  The  fluid  exuded  from  the  foot  of  Polynices  lewisii  was 
also  analyzed.  A  direct  puncture  in  the  dorsal  region  allowed  the 
aspiration  of  body  fluid  from  the  nudibranch  Anisodoria, 

Atnphintura.—  Thp  large  form  Cryptochiton  was  the  only 
r»'pro«ontative  of  this  class  oxaminod.  Fluid  was  obtained  from 
the  pericardial  cavity  by  aspirating  through  a  needle  introduced 
into  the  latter  between  two  adjacent  valves. 

Cyclostomata. 

The  lamprey  Entosphenus  tridentatus  was  obtained  while 
ascending  a  creek  to  spawn.  Blood  was  obtained  by  severing 
the  dorsal  aorta  posterior  to  the  last  gill  slit.  The  blood  was 
oxalatod  as  collected. 

Pisces. 

Blood  was  collected  from  various  specimens  by  severing  the 
caudal  vessels.  It  was  oxalated  as  obtained  and  examined  at 
once. 

Reptilia. 

Two  garter  snakes  were  caught  while  feeding  at  the  water's 
edge.  They  were  bled  from  the  caudal  artery.  The  analysis  of 
this  blood  was  made  for  comparative  purposes  only. 

RKSILTS  AND  DISCUSSION. 


The  results  of  the  analyses  of  the  various  samples  are  shown 
in  Table  I. 

As  might  be  anticipated  the  carbon  dioxide  content  of  the 
blood  or  celomic  fluid  of  marine  forms  is  relatively  ver}-  low  as 
compared  with  the  blood  of  mammals.  The  carbon  dioxide  con- 
tent of  sea  water  in  the  region  from  which  the  specimens  were 


J.  B.  Collip 


333 


secured  is  of  a  widely  varying  quantity.  The  influx  .r  ..  inrge 
volume  of  fresh  water  emanating  from  the  water-sheds  of  the 
coast  ranges,  the  varj'ing  pflfppts  of  tides,  winds,  and  currrents  all 
contribute  towards  a  constantly  changing  density  of  the  sea  water 
in  this  vicinity.  In  Table  II  is  shown  the  density  corrected  to 
15°C.  of  sea  v.-ater  collected  daily  from  the  surface  off  the  landing 
stage  of  the  Marine  Station,  Departure  Bay,  for  a  consecutive 
period  of  33  days,  also  the  density  of  depth  samples  taken  as 
indicated  by  means  of  the  Nanson  water  bottle.  There  are  also 
shown  the  temperature  and  the  alkalinity  of  the  vorious  samples 
in  terms  of  cubic  centimeters  of  0.01  n  sodium  hydroxide  required 
just  to  discharge  the  pink  color  imparted  to  100  cc.  of  sea  water 
by  phenolphthalein  and  the  buffer  value  of  Sorenscn  (3)  or  the 
reactivity  of  Mooro  and  Wilson  (4).  This  latter  factor  is  the 
amount  of  0.01  n  acid  required  to  carry  100  cc.  of  sea  water  from 
the  phenolphthalein  to  the  methyl  orange  point.  Moore,  Pri- 
deaux,  and  Herdman  (5)  found  that  the  reactivity  of  sea  water 
taken  at  Port  Krin  did  not  show  seasonal  variation.  It  is  evi- 
dent, however,  an  the  titration  figures  indicate  that  the  buffer 
value  of  sea  water  in  the  vicinity  of  Departure  Bay  varies  directly 
as  the  density.  Moore  and  coworkers  (5)  state  that  this  buffer 
effect  is  due  in  the  main  to  dissolved  magnesium  bicarbonate  and 
not  calcium  bicarbonate  as  usually  stated.  The  slight  variation 
of  the  alkalinity  of  the  water  from  day  to  day  is  of  interest.  It 
is  probably  a.s.soriated,  as  Moore,  Prideaux,  and  Herdman  (.'>) 
have  pointed  out,  with  the  varying  intensity  of  pholcsynthe.sis 
by  microscopic  organisms.  Surface  and  depth  tciw.s  made  daily 
during  the  summer  months  by  Mouncc'  showed  great  varia- 
tion in  the  relative  amounts,  and  in  the  distribution  of  diatoms. 
The  falling  off  of  the  degree  of  alkalinity  of  .sea  water  with  incireas- 
ing  depth  is  suggestive  in  this  connection.  Sorensen  (3)  found 
that  deep  sea  water  was  less  alkaline  than  that  at  the  surface. 
The  total  carbon  dioxide  content  of  surface  samples  of  sea  water 
obtained  in  Departure  Bay  during  the  summer  of  1920  was 
found  to  vary  between  2  and  4  volumes  per  cent,  the  amount 
varying  directly  with  the  specific  gravity.  It  is  therefore  of 
interest  to  find  that  the  amount  of  carbon  dioxide  in  the  blood  or 


'Unpublished  results  quoted  by  courtesy  of  Miss  Irene  Mounce. 


334 


Alkali  Reserve 


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TABLE  II. 


Date. 

.Sourre  of  sample. 

Temper- 
ature. 

Deiuity 
at   S'C. 

Alkalin- 
ity. 

Reactiv- 
ity. 

tuo 

•c. 

ee.  O.Ol  M 

ee.  0.01  N 

NaOH 

H,SO* 

June  27 

Off  landing  stage  at  station. 

169 

1  0178 

1.0 

17.5 

"    28 

*< 

17.8 

1.0146 

11 

16.0 

"    29 

« 

19.5 

1.0140 

1.3 

13.8 

"    30 

M 

18  0 

1.0159 

1.0 

15.7 

July    1 

U 

18.2 

1  0161 

0.7 

14.8 

"      2 

« 

18.0 

1  0167 

13 

15  1 

"      3 

« 

176 

1.0186 

12 

16  5 

"       4 

<l 

176 

1 .0181 

14 

13.9 

"      5 

« 

17.3 

1.0187 

1.7 

16.6 

"      6 

M 

18.0 

1  0196 

15 

17.5 

"      7 

U 

188 

1.0198 

16 

174 

"      8 

u 

20.5 

1.0112 

13 

13.0 

"      9 

u 

20.5 

1.0122 

1.2 

13  5 

"     1(» 

u 

195 

1.0133 

14 

13.8 

"     11 

« 

17.9 

1  0187 

10 

17.0 

"     12 

w 

15.1 

1  0191 

15 

17  5 

"     13 

u 

14  7 

1  0208 

0.4 

19  0 

"     14 

» 

17  2 

1  0094 

0  5 

12  0 

"     15 

u 

19  7 

1.0108 

0.7 

125 

"     IG 

u 

199 

1.0097 

10 

12  3 

"     17 

tt 

21  0 

1  0133 

0.7 

13  6 

"     18 

« 

199 

1.0142 

10 

13  5 

"     19 

M 

17  0 

1  0187 

0.7 

17  2 

"    20 

« 

17  3 

1.0199 

1.7 

16  5 

"    21 

« 

17.1 

1.0192 

13 

16.7 

"     '22 

M 

18  5 

1.0088 

03 

11  3 

"     23 

« 

17.4 

1.0114 

13 

13.0 

"     24 

U 

16.1 

1  0150 

1.5 

147 

"     25 

« 

17.0 

1  0142 

1.5 

14  7 

"     26 

a 

18.2 

1  0119 

13 

13  5 

"     27 

u 

19  3 

1.0115 

10 

13  0 

"     28 

M 

17.8 

1.0115 

10 

13  0 

"     '29 

u 

18.7 

1  orio 

1.0 

13.7 

"       5 

Surface     sample     off     Five 
Fingers. 

17  2 

1  0203 

16 

19  0 

"       5 

At     5     fathoms     off     Five 
Fingers. 

15.4 

V0215 

1.5 

19  0 

"       5 

At     10    fathoms    off     Five 
Fingers. 

10.00 

1  0225 

0  2 

20.5 

"      5 

At    20     fathoms     off     Five 
Fingers. 

8  98 

1.0252 

0.1 

20  8 

340 


Alkali  Reserve 


TAHI.K  II— Cinicludd. 


Ont 

c. 

Hoiircc  of  Bttinplc. 

Temppr- 
atiirp. 

'C. 

Oeniity 
at  15"C 

Alkalin- 
ity. 

Konrtiv- 
ity. 

I3M 

i-r.OOl  N 
SttOII 

CC   0.01  N 

July 

5 

At     .W     fafhdriis     ..IT     Five 

7  92 

1   02«)7 

.,. 

21  0 

ti 

Ti 

At     UK)    fath.iiiii^    ..(T     Five 
Fingers. 

S  01 

1  0207 

-0.1 

21.0 

it 

12 

At    I   fathiitn  ulT   D('j)artun' 

liiiy. 

14  3 

1  ()21-t 

2  0 

18  5 

ii 

12 

At  2  fathoms  (il'f  I).>parturp 
H;iy. 

1  ()217 

15 

180 

U 

12 

At  .'{  fatlioiiis  ('tT  Departure 
Hay. 

1  0220 

14 

18  5 

ii 

12 

.\t  !>  fathoms  niT  Departure 
Hay. 

1  ()222 

10 

IS  6 

ti 

12 

.\t  M)  fathiinis  olV  Departure 
Hay. 

1  ()2;« 

-0  3 

21  S 

n 

12 

.\t  20  fathoms  otV  Departure 

1  025;- 

-14 

24  0 

I5av. 

fcloiiiic  fluid  of  tho  various  marine  fitriu.s  invest ij^atcd  is  invari- 
ably groat  or  than  llio  niaxitnuni  valuo  for  total  oarbon  dioxido  in 
soa  water.  '['\u)  amount  of  oombinod  carbon  dioxide  in  the  body 
ti.ssuos  of  Modusie  and  soa  anomonos.  the  colomic  fluid  of  brach- 
iopod.«,  .starfish,  certain  soa  urchins,  and  a  number  of  mollusks 
and  tho  blood  of  the  dogfi.sh  and  the  ratfish  i.s  relatively  low,  but 
in  all  instances  is  higiior  than  that  in  sea  water.  Certain  types 
of  C'ru.stacea  h.ave  a  relatively  hijih  carbon  dioxide  content,  such 
forms  as  (\tNci r  iiKtijista-,  ('afic r  /n'otliiclux,  Kchidnoccru^forinalnx, 
and  III  iiHiirtipxnK  nmlns  boing  included  in  this  group.  The  kelp 
oral.)  Kpinltus  produrlh-^,  the  sand  shrimp  CpD'jchin  pimettcnsis, 
and  the  (>;iglc'  l.arnacio  Uiil'unis  lupiiUn  are  on  the  contrary  (!om- 
parativoly  low  in  the  fi.vod  carbon  dioxich.'  in  the  celomic  Huid 
but  they  arc  in  tlii.s  respect  somewhat  analogous  with  the  toleosts 
examined. 

.\  sotiiewlial  .similar  phenomenon  oxi.sts  in  the  case  of  tho 
Molhisca.  While  the  majority  of  the  forms  studied  have  a 
carbon  flioxide  factor  falling  within  the  range  between  6.3  and 
11  volumes  i)er  cent  certain  species  such  as  Puphia  slamfmu, 
and  I'mitilht  pinitu  of  tho  pelocypod  type  and  Polynias  lewisii 


ramn^BSwaaufai 


si.'!dBaKi&;;saESB3K^scr 


J.  B.  CoUip 


341 


and  Thais  lamellosa  of  the  gastropod  class  have  a  fixed  carbon 
dioxide  content  which  in  nearly  all  instances  is  considerably 
higher  than  that  observed  in  the  former  group. 

As  pointed  out  earUer  in  the  paper  it  is  essential  that  the  blood 
be  obtained  while  the  animal  is  perfectly  fresh.  For  instance  it 
was  noted  that  specimens  of  Mya  arenaria  bled  immediately 
after  they  had  been  dug  gave  a  fixed  carbon  dioxide  factor  of  6.5 
volumes  per  cent  while  other  specimens  carried  to  the  laboratory 
in  fresh  sea  water  gave  a  factor  of  8.3  volumes  per  cent  for  com- 
bined carbon  dioxide.  The  fixed  carbon  dioxide  of  the  mollusk 
tends  to  rise  rapidly  when  the  animal  is  not  kept  in  a  large  vol- 
ume of  fresh  water  while  the  opposite  effect  was  noted  in  the  case 
of  the  fish  examined.  Exposure  to  air  causes  the  carbon  dioxide 
content  to  fall  while  in  the  dead  fish  the  latter  may  be  near  to 
that  of  .sea  water.  If  a  fish  is  allowed  to  remain  hooked  but  left 
in  tiic  open  water  for  some  little  time  the  carbon  dioxide  content 
of  the  blood  falls.  The  fi.sh  are  in  their  reaction  to  injurv  much 
hke  the  mammal.-  as  far  as  the  alkali  reserve  of  the  blood  is 
concerned. 

'J-he  relatively  low  figure  for  carbon  dioxide  in  the  blood  of  the 
elasmobranch  Sfjiialus  sucklii  and  the  holocephalan  Hydroktgux 
colliei  stand.s  in  .sharp  contrast  with  that  for  the  teleostian  types 
studied. 

As  the  hydrogen  ion  concentration  of  sea  water  is  in  most 
in.stances  lower  than  that  obtaining  in  the  blood  of  marine  forms 
and  as  the  bicarbonate  content  of  the  latter  is  much  higher  than 
that  of  the  former  it  is  evident  that  the  amount  of  the  dissolved 
carbon  dioxide  in  the  blood  or  body  fluids  of  marine  forms  must 
be  considerably  greater  than  that  occurring  in  sea  water.  The 
tension  of  carbon  dioxide  in  the  blood  of  marine  forms  must 
al.so  he  jjioimrtionately  higher  than  that  in  .sea  water.  This 
brings  up  an  interesting  point.  Does  a  process  of  simple  diffusion 
furnish  an  adequate  explanation  of  the  mode  of  elimination  of 
carbon  dioxide  from  the  blood  or  body  fluids  of  marine  forms 
to  the  surrounding  sea  water?  The  maintenance  of  a  definite 
acid-base  balance  in  the  blood  and  tissues  of  marine  forms  is  no 
doubt  quite  as  prime  an  essential  as  in.  the  case  of  higher  forms. 
In  the  mammal  this  is  largely  effected  by  the  concerted  action  of 
the   respiratory   and   renal   apparatus.     The   gaseous   exchange 


JSBSS'S^St-iii^ 


342 


Alkali  Reserve 


botwpon  tho  blood  and  the  atmosphere  takes  place  almost  entirely 
in  the  alvpolar  sacks  where  the  tensions  of  the  gases  concerned 
arc  such  that  a  process  of  physical  diffusions  seems  to  furnish  a 
sufficient  explanation  for  the  passage  of  the  oxygen  inward  and 
of  the  carbon  dioxide  outward  (G).  In  the  case  of  the  fish  con- 
ditions are  somewhat  different.  While  tho  respiratory  and  renal 
apparatus  here  as  in  mammals  is  apparently  closely  associated 
with  the  regulation  of  the  reaction  of  the  blood  yet  the  inter- 
change of  gases  between  the  blood  and  sea  water  taking  place  as 
it  does  largely  through  the  medium  of  the  gill  filaments  is  on  a 
somewhat  different  basis  from  the  exchange  in  the  mammal  at 
the  lung  surface.  The  tension  of  dissolved  gases  in  sea  water  to 
which  the  gill  filaments  are  exposed  cannot  differ  greatly  from 
the  tensions  obtaining  in  the  enveloping  medium,  unle.ss  it  is 
possible  that  the  respiratory  movements  an;  so  adjuste<l  that  the 
tension  of  gases  in  the  water  of  the  gill  cavities  as  kept  in  equi- 
Jibriuiii  with  the  tension  of  gases  in  the  blood,  in  which  case  a 
process  of  physical  diffusion  would  be  a  sufficient  explanation  of 
the  mode  of  ga.seous  exchange  in  the  fish.  That  this  latter  condi- 
tion should  hold  seems  plausible.  It  may  also  be  possible  that  the 
|)ernieability  of  the  gill  membranes  to  carbon  dioxide  is  such  as  to 
allow  a  steej)  pi'cssure  gradient  to  exist  between  the  dissolved 
carbon  dioxi(l(>  in  the  blood  on  the  one  .side,  and  in  the  sea  water 
on  the  other.  I'urthcr  experiment  only  can  furnish  a  full  expla- 
nation of  this  phenomenon. 

In  the  case  of  arthro{)od  types,  such  as  Cnnctr  mofiister,  Echid- 
noarus  format iiy  nd  others,  the  combined  carbon  dioxide  is  at  a 
much  higher  concentration  than  it  is  in  any  of  the  Pisces  exam- 
ined. ,\  relatively  greater  difference  must  therefore  exist  be- 
tween the  tension  of  carbon  dioxide  in  the  blood  of  these  forms 
and  sea  water  than  in  the  case  of  the  Teleostei.  If  one  is  to 
ex|)lain  the  mode  of  carbon  dioxide  excretion  in  these  forms  by  a 
process  of  physical  diffusion  one  must  assume  that  the  perme- 
ability of  the  gill  filaments  to  carbon  dioxide  is  of  a  very  low  order 
allowing  a  very  steep  pressure  gradient  to  be  maintained  between 
the  two  sides  of  the  medium  for  gaseous  exchange.  Certain  of 
the  Mollusca  have  a  relatively  high  concentration  of  bicarbonate 
in  the  body  fluids  but  it  may  possibly  be  due  to  the  anatomical 
features  occurring  here  for  the  carbon  dioxide  in  the  sea  water 


tn:«ii» 


J.  B.  CoUip 


343 


which  is  bathing  the  organisms  to  exist  at  a  higher  tension  than 
in  the  open  sea. 

Such  a  difference  does  not  exist  Ijetwceii  the  hicurhonate  con- 
tent of  the  body  fluids  and  tissues  of  Echinodermata,  Brachi- 
poda,  and  Ccelenterata,  and  that  of  sea  water  as  hjis  been  noted 
in  the  forms  above  mentioned.  The  existence,  however,  of  a 
definite  pressure  gradient  for  carbon  (Uoxide  Iwtwcen  the  tissue 
and  the  sea  water  suggests  that  a  definite  mechanism  exists  for 
regulating  the  tension  of  this  gas  in  the  body  fluids  and  tissues. 

.SI'MMAIIV. 

1.  The  cjirboh  dioxide  (•(Mitfiil  of  ||m>  blood  :iiid  llic  celornic 
(liiitls  of  various  m.-irinr  forms  has  bn-ri  ilcdTinincd. 

2.  Tlic  cjirlHin  dioxidf  coiitciil  of  the  blood  and  (•(•ioiiii(^  lliiid 
of  marine  forms  cxamiiicil  ((luilibralcd  with  atmosplicrii-  air  is  in 
all  iiistaiKrcs  higher  than  the  carbon  di((xid(>  content  of  sea  water. 

'.i.  The  carbon  dioxide  content  of  certain  of  the  Artlirop«Kla 
and  Mollusca  is  relatively  very  high. 

4.  The  carbon  dioxide  content  of  the  blood  of  marine  Teleostei 
is  approximately  10  volunics  jwr  cent. 

.*>.  The  carbon  dioxide  content  of  the  elasmobranch  Siinalas 
liucklii  and  the  liolocef)h;dan  Hydrohnjm  collici  is  relatively  very 
low. 

0.  The  alkalinity  and  the  reactivity  of  several  samples  of  sea 
water  have  been  determined. 

7.  Surface  .samples  of  .sea  water  in  the  vicinity  of  Departure 
Bay  are  invariably  alkaline  to  phenolphthalein. 

8.  I(  is  held  tiiat  in  ord(>r  to  maintain  the  constant  reaction  of 
th(>  bloo<l  or  body  fluids  of  marine  forms  the  carbon  dioxide 
tension  nm.xt  b»>  considerably  higher  in  the  blood  .and  body  fluids 
than  it  is  in  se.i  water. 

n.  The  (ineslion  of  carixtn  dioxide  excretion  is  tliscu.s.sed. 

In  conclusion  I  desire  to  express  my  thanks  to  the  Curator  of 
the  Biological  Station  at  Departure  Bay,  Dr.  C.  INFacLean  Fraser, 
for  his  kind  assistance  in  making  the  collection  of  material  pos- 
sible, and  for  aid  in  the  identification  of  specimens.  ^My  thanks 
are  also  due  to  the  Biological  Board  of  Canada  for  defraying  the 
expenses  in  connection  with  this  investigation. 


ii^j^ir.^£i4»-."  it.i'^i^l&iH 


344 


Alkali  Reserve 


BIBLIOORAPHY. 

1.  Van  Slyke,  D.  D.,  J.  Biol.  Chem.,  1917,  xxx,  »17. 

•-».  Van  Slyke.  D.  D.,  and  CuUen,  G.  E.,  J.  Biol.  Chem.,  1917,  xxx,  289. 

3.  S<irensen,  S.  P.  F<.,  in  Moore,  Prideatix,  and  Hcrdman  (5),  p.  174. 

4.  Moore,  B.,  and  Wilson,  F.  P.,  Biochem.  J.,  1906,  i,  •Ji?. 

5.  Moore,  H.,  Pridcaux,  E.,  and  Herdman,  C,  Tr.  Biol.  Soc.  Liverpool, 

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6.  Krogh,  .\.,  Skand.  Arch.  Physiol.,  1910,  xxiii,  274. 


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